Characterization of purified myocilin : glaucoma as a protein misfolding disease DEIA Supplement

纯化肌纤蛋白的表征：青光眼作为一种蛋白质错误折叠疾病 DEIA 补充

• 批准号: 10789112
• 负责人: Raquel L Lieberman
• 金额: $ 38.46万
• 依托单位: GEORGIA INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-03-01 至 2026-02-28
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10789112
• 关键词: Academia; Acceleration; Affect; Alleles; Alzheimer' s Disease; Amyloid; Amyloidosis; Amyotrophic Lateral Sclerosis; Anatomy; Anterior eyeball segment structure; Aqueous Humor; Blindness; Cell Death; Cells; Comprehension; Deuterium; Diagnosis; Disease; Drainage procedure; Endoplasmic Reticulum; Extracellular Matrix; Extracellular Matrix Proteins; Eye; Eye diseases; Foundations; Functional disorder; Funding; Glaucoma; Goals; Homeostasis; Horns; Hydrogen; Industry; Inherited; Knowledge; Lead; Length; Liquid substance; Mass Spectrum Analysis; Medical; Methods; Modality; Molecular; Molecular Chaperones; Mutation; Nerve Degeneration; Nuclear Magnetic Resonance; Open-Angle Glaucoma; Outcome; Pathogenesis; Pathogenicity; Pathway interactions; Patients; Physiologic Intraocular Pressure; Physiology; Play; Process; Proteins; Relaxation; Retina; Role; Structure; Therapeutic; Tissues; Trabecular meshwork structure; Variant; amyloid formation; blind; cytotoxicity; early onset; gain of function; infancy; multidisciplinary; mutant; myocilin; novel; novel therapeutics; olfactomedin; pressure; protein misfolding; proteostasis; solid state nuclear magnetic resonance; superoxide dismutase 1; symptom treatment; timeline

Glaucoma, a leading cause of blindness worldwide (70 million patients), is managed medically by treating the symptom of increased intraocular pressure (IOP), but 10% of patients still go blind. IOP is controlled in the anterior segment of the eye, which contains the trabecular meshwork (TM) extracellular matrix, the anatomical pathway for drainage of aqueous humor fluid. The TM tissue is diseased in most forms of glaucoma; loss of TM homeostasis leads to elevated IOP. Hereditary open angle glaucoma, affecting ~3 million young patients, is caused by mutations in myocilin, a protein highly expressed in the TM. Since 3/2011, studies funded from R01EY021205 have changed the paradigm for anti-glaucoma therapeutics by laying the molecular foundation for approaches that target the disease process, which are now being pursued in academia and industry. Studies from R01EY021205 have clarified molecular details of the toxic gain-of-function pathogenic mechanism in which mutant myocilin accumulates in the endoplasmic reticulum (ER) of TM cells, leading to TM cell death and an accelerated timeline for vision loss. Studies from R01EY021205 (a) contributed fundamental knowledge of myocilin structure, (b) discovered a counter-productive interaction between myocilin and the ER-resident Hsp90 chaperone Grp94, and (c) characterized myocilin misfolding as amyloid. Wild-type and many different myocilin variants harbor a misfolding propensity; thus, proteostasis issues identified in familial myocilin-associated glaucoma are likely at play in many more patients. Amyloid formation by myocilin places glaucoma alongside more well-studied amyloid diseases like Alzheimer and SOD-1 dependent amyotrophic lateral sclerosis, yet comprehension of the role of amyloid in glaucoma is in its infancy. The current objective is to better understand molecular aspects of myocilin fibrilization, focused on the relevant olfactomedin (OLF) domain. The multidisciplinary team led by Raquel Lieberman will (a) clarify initiation of aggregation by studying solution structures of wild-type and selected OLF variants, as well as corresponding multi-length scale dynamics, using hydrogen-deuterium exchange mass spectrometry and nuclear magnetic resonance (NMR) structure and relaxation methods (Wade Van Horn, Co- I) (b) compare the end-point structures of selected OLF aggregates to known amyloids by solid state NMR (Anant Paravastu, Co-I) and evaluate cytotoxicity of intermediate aggregates and (c) evaluate common allele full-length myocilin variants for experimental hallmarks of pathogenicity. The expected outcome is a better understanding of the myocilin misfolding process at the molecular level, including molecular determinants of pathogenicity, to enable novel modalities for studying, diagnosing, and treating myocilin-associated glaucoma. More broadly, continued structure/dysfunction studies of myocilin will not only contribute to an understanding of glaucoma and its role in the TM, but will also extend comprehension of the many other OLF domains, which are implicated broadly in physiology and diseases.

青光眼是全球失明的主要原因（7,000万患者），通过治疗来管理 眼内压力增加（IOP）的症状，但10％的患者仍然失明。 IOP在 眼睛的前部，其中包含小梁网（TM）细胞外基质，解剖学 流体流体排水的途径。 TM组织以大多数形式的青光眼患病。损失 TM稳态导致IOP升高。遗传性开倾角青年瘤，影响约30万年轻患者， 是由在TM中高度表达的肌动蛋白突变引起的。自3/2011以来，从 R01EY021205通过奠定分子基础改变了抗云可瘤疗法的范例 对于针对疾病过程的方法，该方法现在正在学术界和工业中采用。 来自R01EY021205的研究阐明了功能性毒性致病性的分子细节 突变体肌动蛋白在TM细胞的内质网（ER）中积聚的机制，导致 TM细胞死亡和视力丧失的加速时间表。 R01EY021205（a）的研究贡献 肌动蛋白结构的基本知识，（b）发现了肌动蛋白之间的适得其反相互作用 以及ER居住的HSP90伴侣GRP94，（C）将肌动蛋白折叠折叠为淀粉样蛋白。野生型 许多不同的肌动蛋白变体具有错误折叠的倾向。因此，在 在更多患者中，家族性肌动蛋白相关的青光眼可能会发挥作用。 肌动蛋白的淀粉样蛋白形成将青光眼与更精心培养的淀粉样蛋白疾病一样 阿尔茨海默氏症和SOD-1依赖性肌萎缩性横向硬化症，但理解淀粉样蛋白在 青光眼仍处于起步阶段。当前的目标是更好地了解肌动蛋白的分子方面 纤维化，重点是相关的嗅觉素（OLF）结构域。拉奎尔（Raquel）领导的多学科团队 利伯曼（Lieberman 使用氢欧交换质量 光谱和核磁共振（NMR）结构和松弛方法（Wade Van Horn，共同 i）（b）将选定的OLF聚集体的终点结构与固态NMR的已知淀粉样蛋白进行比较 （Anant Paravastu，Co-I）并评估中间聚集体的细胞毒性，并评估公共等位基因 全长肌动蛋白变体，用于致病性实验标志。预期的结果更好 了解分子水平的肌动蛋白错误折叠过程，包括分子决定因素 致病性，以实现研究，诊断和治疗肌动蛋白相关的青光眼的新型方式。 更广泛的是，肌动蛋白的持续结构/功能障碍研究不仅有助于理解 青光眼及其在TM中的作用，但也将扩展对许多其他OLF领域的理解， 广泛涉及生理和疾病。